Method development for separation and quantitative determination of sulfacetamide sodium, betamethasone, methyl paraben and propyl paraben in ophthalmic drops

This study includes the development of spectrophotometric and chromatographic methods for determination of Sulfacetamide sodium(SFS) and Betamethasone(BTM) active agents, Methyl paraben(MP) and Propyl paraben(PP) excipient substance in a pharmaceutical ophthalmic drop. In spectrophotometric method, fourth order derivative at 268 nm n=17 was selected for the determination of SFS, third order derivative of the at 236 nm n=21 was selected for the determination of BTM and third order derivative of the 283 nm n=11 was selected for the determination of together MP and PP. Linear operating range was obtained as 6,0-14,0, 10,0-30,0 ve 4,0-12,0 µg/mL for SFS, BTM, and MP-PP respectively. The LOD values were found to be 0,58; 0,89 and 1,14 and µg/mL for SFS, BTM and MP-PP, in spectrophotometric method, respectively. As chromatographic method, the reverse phase high performance liquid chromatographic (RP- HPLC) method has been developed by using column Agilent Zorbax Eclipse XDB C18 (3.0x75mm, 3.5µm). The mobil phase consisted water and acetonitrile (55:45 v/v), the flow rate was 0.5 ml/min and the UV detector was detected at 250 nm. The linear ranges in the method were 3,0-7,0 µg/mL; 1,5-3,5 µg/mL; 0,6-1,4 µg/mL and 0,6-1,4 µg/mL for SFS, BTM, MP and PP, respectively. The LOD values in the validation studies were 0,37; 0,052; 0,021 and 0,012 µg/mL for SFS, BTM, MP and PP in RP-HPLC method respectively. The results of the chromatographic and spectrophotometric methods developed were statistically compared with each other in terms of accuracy and precision, and there was no significant difference between the two methods.